摘要
【目的】将栽培稻品种恢复为米质优、抗逆性强的红稻具有较大的研究价值。利用CRISPR/Cas9基因编辑技术,编辑原花青素转录调节因子Rc基因,恢复红种皮特性,以改良水稻米质,提升抗逆性。【方法】利用CRISPR/Cas9技术,以Rc为靶基因,构建突变载体pYLCRISPR/Cas9-Rc-gRNA,以空育180、上育453为材料,转化获得转基因植株,通过测序手段和表型观察验证成果。【结果】分子水平检测获得Rc突变材料2种,其中KY-1在1414―1417 bp缺失4个碱基,终止子突变为苯丙氨酸;SY-1在1411 bp处缺失1个碱基,终止子突变为天冬氨酸。2种编辑材料均恢复为红米表型,且具有一定耐盐碱能力。【结论】利用CRISPR/Cas9基因编辑技术成功获得恢复红种皮表型的纯合株系,为红米改良提供基础材料。
【Objective】It is of great value to restore cultivated rice varieties to red rice with good rice quality and strong stress resistance.CRISPR/Cas9 gene editing technology was used to edit the Rc gene,restoring the red seed coat,and laying a data foundation for the improvement of rice quality and resistance.【Methods】Using CRISPR/Cas9 technology,the target gene Rc mutant vector pYLCRISPR/Cas9-Rc-gRNA was constructed and transformed into transgenic plants with Kongyu 180 and Shangyu 453 as materials.The results were verified by sequencing and phenotypic observation.【Results】Two Rc mutants were obtained.For KY-1,with 4 bases deleted from 1414 bp to 1417 bp,the terminator was replaced by phenylalanine.SY-1 also lost a base at 1411 bp,which resulted in the transformation from the terminator to aspartic acid.The two kinds of editing materials restored red rice phenotype and had certain saline-alkali tolerance.【Conclusion】Using CRISPR/Cas9 technique,we obtained homozygous lines with red seed coat,providing basic materials for the improvement of red rice.
作者
张元野
尹丽颖
李荣田
何明良
刘欣欣
潘婷婷
田晓杰
卜庆云
李秀峰
ZHANG Yuanye;YIN Liying;LI Rongtian;HE Mingliang;LIU Xinxin;PAN Tingting;TIAN Xiaojie;BU Qingyun;LI Xiufeng(College of Life Science,Heilongjiang University,Harbin 150080,China;Northeast Institute of Geography and Agroecology,Key Laboratory of Soybean Molecular Design Breeding,Chinese Academy of Sciences,Harbin 150081,China;College of Life Science,Northeast Forestry University,Harbin 150040,China;College of Agriculture,Heilongjiang Bayi Agricultural University,Daqing 163319,China)
出处
《中国水稻科学》
CAS
CSCD
北大核心
2022年第6期572-578,共7页
Chinese Journal of Rice Science
基金
黑龙江省杰出青年基金资助项目(JQ2020C003)
中国科学院战略性先导科技专项(A类)(XDA28100000)。
